BISC 219/F10:Calendars/Planner - Revision history

Tucker Crum: /* Schedule of Experiments */

2010-10-15T16:21:53Z

Schedule of Experiments

←Older revision		Revision as of 16:21, 15 October 2010
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	!2		!2
	\| Classical (Forward) Genetics: Mutant Hunt, Linkage, Mapping, Complementation, DNA Sequence analysis		\| Classical (Forward) Genetics: Mutant Hunt, Linkage, Mapping, Complementation, DNA Sequence analysis
-	\| 2-6	+	\| 2-7
	\|-		\|-
	!3		!3

Tucker Crum: /* BISC219 F10 Weekly Lab Planner */

2010-10-05T14:05:59Z

BISC219 F10 Weekly Lab Planner

←Older revision		Revision as of 14:05, 5 October 2010
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	\| '''Series3: Reverse Genetics''': Plasmid isolation from BL21 cells/quantification of DNA/ transformation of plasmid into HT115(DE3)cells		\| '''Series3: Reverse Genetics''': Plasmid isolation from BL21 cells/quantification of DNA/ transformation of plasmid into HT115(DE3)cells
	\| '''Day before lab:''' grow overnight culture of single colony from transformation		\| '''Day before lab:''' grow overnight culture of single colony from transformation
-	\| '''Homework''': Write all Series3 Protocols as Materials & Methods. Assignment described at [[BISC_219/F10: Assignment_Series3_ Materials and Methods]]. Due at the beginning of Lab 9 ; Read the original 1998 paper by Fire and Mello published in ''Nature'' '''391''': 806-811 (Feb.19 1998) at [http://dx.doi.org/10.1038/35888 doi:10.1038/35888] ; Do your own library search to find and read a recent review article about RNAi in ''C. elegans''. Also read the 2002 Simmer ''et al.'' paper in ''Current Biology'' '''12''':1317-1319 explaining the ''rrf-3'' strain that we will use for our RNAi work at [http://dx.doi.org/10.1016/S0960-9822(02)01041-2 doi:10.1016/S0960-9822(02)01041-2 ]. These articles provide background information on how the mechanism of RNAi was worked out experimentally and will be helpful to you when you write the introduction section of your Series3 paper. Also read a published journal article by Green et al. (2009) Impact of Cigarette Smoke Exposure on Innate Immunity: A ''Caenorhabditis elegans'' Model. PLoS ONE 4(8): e6860, found at [http://dx.doi.org/10.1371/journal.pone.0006860 doi:10.1371/journal.pone.0006860]. This study, like yours, uses reverse genetics to investigate gene function~~. There are questions to guide your reading found at:[[BISC_219:Questions to Guide Your Reading 2]]~~ . Be prepared to discuss ~~those questions~~ in Lab 9.	+	\| '''Homework''': Write all Series3 Protocols as Materials & Methods. Assignment described at [[BISC_219/F10: Assignment_Series3_ Materials and Methods]]. Due at the beginning of Lab 9 ; Read the original 1998 paper by Fire and Mello published in ''Nature'' '''391''': 806-811 (Feb.19 1998) at [http://dx.doi.org/10.1038/35888 doi:10.1038/35888] ; Do your own library search to find and read a recent review article about RNAi in ''C. elegans''. Also read the 2002 Simmer ''et al.'' paper in ''Current Biology'' '''12''':1317-1319 explaining the ''rrf-3'' strain that we will use for our RNAi work at [http://dx.doi.org/10.1016/S0960-9822(02)01041-2 doi:10.1016/S0960-9822(02)01041-2 ]. These articles provide background information on how the mechanism of RNAi was worked out experimentally and will be helpful to you when you write the introduction section of your Series3 paper. Also read a published journal article by Green et al. (2009) Impact of Cigarette Smoke Exposure on Innate Immunity: A ''Caenorhabditis elegans'' Model. PLoS ONE 4(8): e6860, found at [http://dx.doi.org/10.1371/journal.pone.0006860 doi:10.1371/journal.pone.0006860]. This study, like yours, uses reverse genetics to investigate gene function. Be prepared to discuss this paper in Lab 9.

	\|-		\|-

Tucker Crum: /* Links to Labs& Project Info */

2010-10-01T01:48:46Z

Links to Labs& Project Info

←Older revision		Revision as of 01:48, 1 October 2010
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	[[BISC 219/F10: Lab 3 \| Lab 3: Linkage Test Part 1]]<br>		[[BISC 219/F10: Lab 3 \| Lab 3: Linkage Test Part 1]]<br>
	[[BISC 219/F10: Lab 4 \| Lab 4: Linkage Test Part 2, Mapping and Complementation]]<br>		[[BISC 219/F10: Lab 4 \| Lab 4: Linkage Test Part 2, Mapping and Complementation]]<br>
-	[[BISC 219/F10: Lab 5 \| Lab 5: Mapping Con't]]<br>	+	[[BISC 219/F10: Lab 5 \| Lab 5: Finish Complementation; Mapping Con't]]<br>
-	[[BISC 219/F10: Lab 6 \| Lab 6: ~~Finish Complementation~~; Mapping Con't]]<BR>	+	[[BISC 219/F10: Lab 6 \| Lab 6: DNA sequence analysis; Mapping Con't]]<BR>
	[[BISC 219/F10: Lab 7 \| Lab 7: Complete Mapping: Score]]<br>		[[BISC 219/F10: Lab 7 \| Lab 7: Complete Mapping: Score]]<br>
	Series3:<BR>		Series3:<BR>

Tucker Crum: /* BISC219 F10 Weekly Lab Planner */

2010-10-01T01:24:11Z

BISC219 F10 Weekly Lab Planner

←Older revision		Revision as of 01:24, 1 October 2010
Line 108:		Line 108:
	\| '''Series2:Mapping:'''Self double mutants to keep viable true-breeding progeny;<BR>'''Series2:Complementation:''' examine cross plates for Dpy males - WHY?; '''Series3: Reverse Genetics:''' Pick gene of interest; set up PCR reaction to clone the gene		\| '''Series2:Mapping:'''Self double mutants to keep viable true-breeding progeny;<BR>'''Series2:Complementation:''' examine cross plates for Dpy males - WHY?; '''Series3: Reverse Genetics:''' Pick gene of interest; set up PCR reaction to clone the gene
	\| '''Series 3:Reverse Genetics:''' Examine the results of agarose gel electrophoresis		\| '''Series 3:Reverse Genetics:''' Examine the results of agarose gel electrophoresis
-	\| '''Homework:''' Explain complementation analysis in general and specifically how it was used to id~~, potentially anyway,~~ your ''dpy'' gene and to determine if the mutation of interest has been previously characterized. Diagram complementation crosses. Template downloadable at: [[Media:Complementation_Template_Crosses.pptx]]Due at the beginning of Lab 6. Assignment described at[[BISC_219/F10: Assignment_Series2_Complementation]]	+	\| '''Homework:''' Explain complementation analysis in general and specifically how it was used to id your ''dpy'' gene and to determine if the mutation of interest has been previously characterized. Diagram complementation crosses. Template downloadable at: [[Media:Complementation_Template_Crosses.pptx]]Due at the beginning of Lab 6. Assignment described at[[BISC_219/F10: Assignment_Series2_Complementation]]
	\|-		\|-
	! 6		! 6

Tucker Crum: /* BISC219 F10 Weekly Lab Planner */

2010-10-01T01:03:14Z

BISC219 F10 Weekly Lab Planner

←Older revision		Revision as of 01:03, 1 October 2010
Line 113:		Line 113:
	\| Oct. 13 to <br> Oct. 19		\| Oct. 13 to <br> Oct. 19
	\| '''Series 2: Mapping''': cross N2 males (++/++) with L4 double mutant hermaphrodites (2 plates total);Self double mutants to new plate<BR> Series2: Characterizing the dpy mutation''' Gene Sequence Analysis <br> '''Series3: Reverse Genetics:'''Restriction enzyme digest PCR product/clean-up/ligation into pPD129.36 vector/transformation into BL21 ''E.coli''		\| '''Series 2: Mapping''': cross N2 males (++/++) with L4 double mutant hermaphrodites (2 plates total);Self double mutants to new plate<BR> Series2: Characterizing the dpy mutation''' Gene Sequence Analysis <br> '''Series3: Reverse Genetics:'''Restriction enzyme digest PCR product/clean-up/ligation into pPD129.36 vector/transformation into BL21 ''E.coli''
-	\| '''3 days after lab:Series2: Mapping:''' pick (4) male heterozygotes (++/d u) and Cross them with L4 hermaphrodite double mutants (d u/d u). <BR>Series 3:Check control & transformation plates (save) – <BR>notify instructor if no colonies	+	\| '''3 days after lab:Series2: Mapping:''' pick (4) male heterozygotes (++/d u) and Cross them with L4 hermaphrodite double mutants (d u/d u). <BR>'''Series 3:'''Check control & transformation plates (save) – <BR>notify instructor if no colonies
	\| '''Homework''': DNA sequencing Analysis due at the beginning of Lab 7. Assignment description at [[BISC_219/F10: Assignment_Series2_DNA Sequencing]]<BR>		\| '''Homework''': DNA sequencing Analysis due at the beginning of Lab 7. Assignment description at [[BISC_219/F10: Assignment_Series2_DNA Sequencing]]<BR>
	Scientific research report on Series2 Classical (Forward) Genetics Project due week of 11/1 at the beginning of lab. 5%/day late penalty! See assignment directions at: [[BISC 219/F10: Assignment_ Series2_Classical Genetics Paper]]		Scientific research report on Series2 Classical (Forward) Genetics Project due week of 11/1 at the beginning of lab. 5%/day late penalty! See assignment directions at: [[BISC 219/F10: Assignment_ Series2_Classical Genetics Paper]]
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	\| Oct. 20 to <br> Oct. 26		\| Oct. 20 to <br> Oct. 26
	\| '''Series2:Finish Mapping:''' SCORE! Calculate Recombination frequency; <BR>'''Series3:Reverse Genetics''' Colony PCR to look for bacterial transformants		\| '''Series2:Finish Mapping:''' SCORE! Calculate Recombination frequency; <BR>'''Series3:Reverse Genetics''' Colony PCR to look for bacterial transformants
-	\| Determine map distance between your mutant gene <BR>and the known reference mutation'''Day before lab:''' grow overnight culture of positive colony	+	\| Determine map distance between your mutant gene <BR>and the known reference mutation<BR>'''Day before lab:''' Series 3:grow overnight culture of positive colony
	\| '''Homework''': Work on your Series 2: Forward genetics paper due week of Nov 1		\| '''Homework''': Work on your Series 2: Forward genetics paper due week of Nov 1
	\|-		\|-

Tucker Crum: /* BISC219 F10 Weekly Lab Planner */

2010-10-01T01:01:42Z

BISC219 F10 Weekly Lab Planner

←Older revision		Revision as of 01:01, 1 October 2010
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	! 5		! 5
	\| Oct. 5 to<br> Oct. 12		\| Oct. 5 to<br> Oct. 12
-	\| '''Series2:Mapping:'''Self double mutants to keep viable true-breeding progeny<BR>'''Series3: Reverse Genetics:''' Pick gene of interest; set up PCR reaction to clone the gene~~;<BR>'''Series2:Complementation:''' examine cross plates for Dpy males - WHY?;~~	+	\| '''Series2:Mapping:'''Self double mutants to keep viable true-breeding progeny;<BR>'''Series2:Complementation:''' examine cross plates for Dpy males - WHY?; '''Series3: Reverse Genetics:''' Pick gene of interest; set up PCR reaction to clone the gene
	\| '''Series 3:Reverse Genetics:''' Examine the results of agarose gel electrophoresis		\| '''Series 3:Reverse Genetics:''' Examine the results of agarose gel electrophoresis
	\| '''Homework:''' Explain complementation analysis in general and specifically how it was used to id, potentially anyway, your ''dpy'' gene and to determine if the mutation of interest has been previously characterized. Diagram complementation crosses. Template downloadable at: [[Media:Complementation_Template_Crosses.pptx]]Due at the beginning of Lab 6. Assignment described at[[BISC_219/F10: Assignment_Series2_Complementation]]		\| '''Homework:''' Explain complementation analysis in general and specifically how it was used to id, potentially anyway, your ''dpy'' gene and to determine if the mutation of interest has been previously characterized. Diagram complementation crosses. Template downloadable at: [[Media:Complementation_Template_Crosses.pptx]]Due at the beginning of Lab 6. Assignment described at[[BISC_219/F10: Assignment_Series2_Complementation]]
Line 113:		Line 113:
	\| Oct. 13 to <br> Oct. 19		\| Oct. 13 to <br> Oct. 19
	\| '''Series 2: Mapping''': cross N2 males (++/++) with L4 double mutant hermaphrodites (2 plates total);Self double mutants to new plate<BR> Series2: Characterizing the dpy mutation''' Gene Sequence Analysis <br> '''Series3: Reverse Genetics:'''Restriction enzyme digest PCR product/clean-up/ligation into pPD129.36 vector/transformation into BL21 ''E.coli''		\| '''Series 2: Mapping''': cross N2 males (++/++) with L4 double mutant hermaphrodites (2 plates total);Self double mutants to new plate<BR> Series2: Characterizing the dpy mutation''' Gene Sequence Analysis <br> '''Series3: Reverse Genetics:'''Restriction enzyme digest PCR product/clean-up/ligation into pPD129.36 vector/transformation into BL21 ''E.coli''
-	\| '''3 days after lab:~~''''''~~Series2: Mapping:''' pick (4) male heterozygotes (++/d u) and Cross them with L4 hermaphrodite double mutants (d u/d u). <BR>Series 3:Check control & transformation plates (save) – <BR>notify instructor if no colonies	+	\| '''3 days after lab:Series2: Mapping:''' pick (4) male heterozygotes (++/d u) and Cross them with L4 hermaphrodite double mutants (d u/d u). <BR>Series 3:Check control & transformation plates (save) – <BR>notify instructor if no colonies
	\| '''Homework''': DNA sequencing Analysis due at the beginning of Lab 7. Assignment description at [[BISC_219/F10: Assignment_Series2_DNA Sequencing]]<BR>		\| '''Homework''': DNA sequencing Analysis due at the beginning of Lab 7. Assignment description at [[BISC_219/F10: Assignment_Series2_DNA Sequencing]]<BR>
	Scientific research report on Series2 Classical (Forward) Genetics Project due week of 11/1 at the beginning of lab. 5%/day late penalty! See assignment directions at: [[BISC 219/F10: Assignment_ Series2_Classical Genetics Paper]]		Scientific research report on Series2 Classical (Forward) Genetics Project due week of 11/1 at the beginning of lab. 5%/day late penalty! See assignment directions at: [[BISC 219/F10: Assignment_ Series2_Classical Genetics Paper]]

Tucker Crum: /* BISC219 F10 Weekly Lab Planner */

2010-10-01T01:00:00Z

BISC219 F10 Weekly Lab Planner

←Older revision		Revision as of 01:00, 1 October 2010
Line 107:		Line 107:
	\| Oct. 5 to<br> Oct. 12		\| Oct. 5 to<br> Oct. 12
	\| '''Series2:Mapping:'''Self double mutants to keep viable true-breeding progeny<BR>'''Series3: Reverse Genetics:''' Pick gene of interest; set up PCR reaction to clone the gene;<BR>'''Series2:Complementation:''' examine cross plates for Dpy males - WHY?;		\| '''Series2:Mapping:'''Self double mutants to keep viable true-breeding progeny<BR>'''Series3: Reverse Genetics:''' Pick gene of interest; set up PCR reaction to clone the gene;<BR>'''Series2:Complementation:''' examine cross plates for Dpy males - WHY?;
-	\| '''Series 3:Reverse Genetics:''' Examine the results of agarose gel electrophoresis~~<BR>'''3 days after lab:'''~~	+	\| '''Series 3:Reverse Genetics:''' Examine the results of agarose gel electrophoresis
	\| '''Homework:''' Explain complementation analysis in general and specifically how it was used to id, potentially anyway, your ''dpy'' gene and to determine if the mutation of interest has been previously characterized. Diagram complementation crosses. Template downloadable at: [[Media:Complementation_Template_Crosses.pptx]]Due at the beginning of Lab 6. Assignment described at[[BISC_219/F10: Assignment_Series2_Complementation]]		\| '''Homework:''' Explain complementation analysis in general and specifically how it was used to id, potentially anyway, your ''dpy'' gene and to determine if the mutation of interest has been previously characterized. Diagram complementation crosses. Template downloadable at: [[Media:Complementation_Template_Crosses.pptx]]Due at the beginning of Lab 6. Assignment described at[[BISC_219/F10: Assignment_Series2_Complementation]]
	\|-		\|-
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	\| Oct. 13 to <br> Oct. 19		\| Oct. 13 to <br> Oct. 19
	\| '''Series 2: Mapping''': cross N2 males (++/++) with L4 double mutant hermaphrodites (2 plates total);Self double mutants to new plate<BR> Series2: Characterizing the dpy mutation''' Gene Sequence Analysis <br> '''Series3: Reverse Genetics:'''Restriction enzyme digest PCR product/clean-up/ligation into pPD129.36 vector/transformation into BL21 ''E.coli''		\| '''Series 2: Mapping''': cross N2 males (++/++) with L4 double mutant hermaphrodites (2 plates total);Self double mutants to new plate<BR> Series2: Characterizing the dpy mutation''' Gene Sequence Analysis <br> '''Series3: Reverse Genetics:'''Restriction enzyme digest PCR product/clean-up/ligation into pPD129.36 vector/transformation into BL21 ''E.coli''
-	\| '''Series2: Mapping:''' pick (4) male heterozygotes (++/d u) and Cross them with L4 hermaphrodite double mutants (d u/d u). <BR>Series 3:Check control & transformation plates (save) – <BR>notify instructor if no colonies	+	\| '''3 days after lab:''''''Series2: Mapping:''' pick (4) male heterozygotes (++/d u) and Cross them with L4 hermaphrodite double mutants (d u/d u). <BR>Series 3:Check control & transformation plates (save) – <BR>notify instructor if no colonies
	\| '''Homework''': DNA sequencing Analysis due at the beginning of Lab 7. Assignment description at [[BISC_219/F10: Assignment_Series2_DNA Sequencing]]<BR>		\| '''Homework''': DNA sequencing Analysis due at the beginning of Lab 7. Assignment description at [[BISC_219/F10: Assignment_Series2_DNA Sequencing]]<BR>
	Scientific research report on Series2 Classical (Forward) Genetics Project due week of 11/1 at the beginning of lab. 5%/day late penalty! See assignment directions at: [[BISC 219/F10: Assignment_ Series2_Classical Genetics Paper]]		Scientific research report on Series2 Classical (Forward) Genetics Project due week of 11/1 at the beginning of lab. 5%/day late penalty! See assignment directions at: [[BISC 219/F10: Assignment_ Series2_Classical Genetics Paper]]

Tucker Crum: /* BISC219 F10 Weekly Lab Planner */

2010-10-01T00:58:46Z

BISC219 F10 Weekly Lab Planner

←Older revision		Revision as of 00:58, 1 October 2010
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	\| Oct. 5 to<br> Oct. 12		\| Oct. 5 to<br> Oct. 12
	\| '''Series2:Mapping:'''Self double mutants to keep viable true-breeding progeny<BR>'''Series3: Reverse Genetics:''' Pick gene of interest; set up PCR reaction to clone the gene;<BR>'''Series2:Complementation:''' examine cross plates for Dpy males - WHY?;		\| '''Series2:Mapping:'''Self double mutants to keep viable true-breeding progeny<BR>'''Series3: Reverse Genetics:''' Pick gene of interest; set up PCR reaction to clone the gene;<BR>'''Series2:Complementation:''' examine cross plates for Dpy males - WHY?;
-	\| '''Series 3:Reverse Genetics:''' Examine the results of agarose gel electrophoresis<BR>'''3 days after lab:''' ~~'''Series2: Mapping:''' pick (4) <BR>heterozygotes from previous cross. Cross these wild type males (+ +/+ +) X your double mutant (d u/d u).~~	+	\| '''Series 3:Reverse Genetics:''' Examine the results of agarose gel electrophoresis<BR>'''3 days after lab:'''
	\| '''Homework:''' Explain complementation analysis in general and specifically how it was used to id, potentially anyway, your ''dpy'' gene and to determine if the mutation of interest has been previously characterized. Diagram complementation crosses. Template downloadable at: [[Media:Complementation_Template_Crosses.pptx]]Due at the beginning of Lab 6. Assignment described at[[BISC_219/F10: Assignment_Series2_Complementation]]		\| '''Homework:''' Explain complementation analysis in general and specifically how it was used to id, potentially anyway, your ''dpy'' gene and to determine if the mutation of interest has been previously characterized. Diagram complementation crosses. Template downloadable at: [[Media:Complementation_Template_Crosses.pptx]]Due at the beginning of Lab 6. Assignment described at[[BISC_219/F10: Assignment_Series2_Complementation]]
	\|-		\|-
	! 6		! 6
	\| Oct. 13 to <br> Oct. 19		\| Oct. 13 to <br> Oct. 19
-	\| '''Series 2: Mapping''': cross N2 males with double ~~mutants~~ (2 plates total);Self double mutants to new plate<BR> Series2: Characterizing the dpy mutation''' Gene Sequence Analysis <br> '''Series3: Reverse Genetics:'''Restriction enzyme digest PCR product/clean-up/ligation into pPD129.36 vector/transformation into BL21 ''E.coli''	+	\| '''Series 2: Mapping''': cross N2 males (++/++) with L4 double mutant hermaphrodites (2 plates total);Self double mutants to new plate<BR> Series2: Characterizing the dpy mutation''' Gene Sequence Analysis <br> '''Series3: Reverse Genetics:'''Restriction enzyme digest PCR product/clean-up/ligation into pPD129.36 vector/transformation into BL21 ''E.coli''
-	\| ~~Series 2~~:~~Cross heterozygous~~ (++/~~dyp unc~~) ~~males~~ with L4 double ~~mutant hermaphrodites~~.<BR>Series 3:Check control & transformation plates (save) – <BR>notify instructor if no colonies	+	\| '''Series2: Mapping:''' pick (4) male heterozygotes (++/d u) and Cross them with L4 hermaphrodite double mutants (d u/d u). <BR>Series 3:Check control & transformation plates (save) – <BR>notify instructor if no colonies
	\| '''Homework''': DNA sequencing Analysis due at the beginning of Lab 7. Assignment description at [[BISC_219/F10: Assignment_Series2_DNA Sequencing]]<BR>		\| '''Homework''': DNA sequencing Analysis due at the beginning of Lab 7. Assignment description at [[BISC_219/F10: Assignment_Series2_DNA Sequencing]]<BR>
	Scientific research report on Series2 Classical (Forward) Genetics Project due week of 11/1 at the beginning of lab. 5%/day late penalty! See assignment directions at: [[BISC 219/F10: Assignment_ Series2_Classical Genetics Paper]]		Scientific research report on Series2 Classical (Forward) Genetics Project due week of 11/1 at the beginning of lab. 5%/day late penalty! See assignment directions at: [[BISC 219/F10: Assignment_ Series2_Classical Genetics Paper]]

Tucker Crum: /* BISC219 F10 Weekly Lab Planner */

2010-10-01T00:53:34Z

BISC219 F10 Weekly Lab Planner

←Older revision		Revision as of 00:53, 1 October 2010
Line 106:		Line 106:
	! 5		! 5
	\| Oct. 5 to<br> Oct. 12		\| Oct. 5 to<br> Oct. 12
-	\| '''Series2:Mapping:Self double mutants to keep viable true-breeding progeny<BR>'''Series3: Reverse Genetics:''' Pick gene of interest; set up PCR reaction to clone the gene;<BR>'''Series2:Complementation:''' examine cross plates for Dpy males - WHY?;	+	\| '''Series2:Mapping:'''Self double mutants to keep viable true-breeding progeny<BR>'''Series3: Reverse Genetics:''' Pick gene of interest; set up PCR reaction to clone the gene;<BR>'''Series2:Complementation:''' examine cross plates for Dpy males - WHY?;
	\| '''Series 3:Reverse Genetics:''' Examine the results of agarose gel electrophoresis<BR>'''3 days after lab:''' '''Series2: Mapping:''' pick (4) <BR>heterozygotes from previous cross. Cross these wild type males (+ +/+ +) X your double mutant (d u/d u).		\| '''Series 3:Reverse Genetics:''' Examine the results of agarose gel electrophoresis<BR>'''3 days after lab:''' '''Series2: Mapping:''' pick (4) <BR>heterozygotes from previous cross. Cross these wild type males (+ +/+ +) X your double mutant (d u/d u).
	\| '''Homework:''' Explain complementation analysis in general and specifically how it was used to id, potentially anyway, your ''dpy'' gene and to determine if the mutation of interest has been previously characterized. Diagram complementation crosses. Template downloadable at: [[Media:Complementation_Template_Crosses.pptx]]Due at the beginning of Lab 6. Assignment described at[[BISC_219/F10: Assignment_Series2_Complementation]]		\| '''Homework:''' Explain complementation analysis in general and specifically how it was used to id, potentially anyway, your ''dpy'' gene and to determine if the mutation of interest has been previously characterized. Diagram complementation crosses. Template downloadable at: [[Media:Complementation_Template_Crosses.pptx]]Due at the beginning of Lab 6. Assignment described at[[BISC_219/F10: Assignment_Series2_Complementation]]

Tucker Crum: /* BISC219 F10 Weekly Lab Planner */

2010-10-01T00:52:50Z

BISC219 F10 Weekly Lab Planner

←Older revision		Revision as of 00:52, 1 October 2010
Line 106:		Line 106:
	! 5		! 5
	\| Oct. 5 to<br> Oct. 12		\| Oct. 5 to<br> Oct. 12
-	\| '''~~Series 2~~: Mapping~~'''~~: ~~cross N2 males with~~ double mutants ~~(2 plates total);<BR>~~ <BR> '''Series3: Reverse Genetics:''' Pick gene of interest; set up PCR reaction to clone the gene;<BR>'''Series2:Complementation:''' examine cross plates for Dpy males - WHY?;	+	\| '''Series2:Mapping:Self double mutants to keep viable true-breeding progeny<BR>'''Series3: Reverse Genetics:''' Pick gene of interest; set up PCR reaction to clone the gene;<BR>'''Series2:Complementation:''' examine cross plates for Dpy males - WHY?;
	\| '''Series 3:Reverse Genetics:''' Examine the results of agarose gel electrophoresis<BR>'''3 days after lab:''' '''Series2: Mapping:''' pick (4) <BR>heterozygotes from previous cross. Cross these wild type males (+ +/+ +) X your double mutant (d u/d u).		\| '''Series 3:Reverse Genetics:''' Examine the results of agarose gel electrophoresis<BR>'''3 days after lab:''' '''Series2: Mapping:''' pick (4) <BR>heterozygotes from previous cross. Cross these wild type males (+ +/+ +) X your double mutant (d u/d u).
	\| '''Homework:''' Explain complementation analysis in general and specifically how it was used to id, potentially anyway, your ''dpy'' gene and to determine if the mutation of interest has been previously characterized. Diagram complementation crosses. Template downloadable at: [[Media:Complementation_Template_Crosses.pptx]]Due at the beginning of Lab 6. Assignment described at[[BISC_219/F10: Assignment_Series2_Complementation]]		\| '''Homework:''' Explain complementation analysis in general and specifically how it was used to id, potentially anyway, your ''dpy'' gene and to determine if the mutation of interest has been previously characterized. Diagram complementation crosses. Template downloadable at: [[Media:Complementation_Template_Crosses.pptx]]Due at the beginning of Lab 6. Assignment described at[[BISC_219/F10: Assignment_Series2_Complementation]]
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	! 6		! 6
	\| Oct. 13 to <br> Oct. 19		\| Oct. 13 to <br> Oct. 19
-	\| '''~~Series2~~:Mapping:''' ~~SCORE!~~; <BR> ~~'''~~Series2:~~Mapping:~~''' Gene Sequence Analysis <br> '''Series3: Reverse Genetics:'''Restriction enzyme digest PCR product/clean-up/ligation into pPD129.36 vector/transformation into BL21 ''E.coli''	+	\| '''Series 2: Mapping''': cross N2 males with double mutants (2 plates total);Self double mutants to new plate<BR> Series2: Characterizing the dpy mutation''' Gene Sequence Analysis <br> '''Series3: Reverse Genetics:'''Restriction enzyme digest PCR product/clean-up/ligation into pPD129.36 vector/transformation into BL21 ''E.coli''
-	\| ~~Determine map distance between your~~ mutant ~~gene <BR>and the known reference mutation~~. <BR>Check control & transformation plates (save) – <BR>notify instructor if no colonies	+	\| Series 2:Cross heterozygous (++/dyp unc) males with L4 double mutant hermaphrodites.<BR>Series 3:Check control & transformation plates (save) – <BR>notify instructor if no colonies
	\| '''Homework''': DNA sequencing Analysis due at the beginning of Lab 7. Assignment description at [[BISC_219/F10: Assignment_Series2_DNA Sequencing]]<BR>		\| '''Homework''': DNA sequencing Analysis due at the beginning of Lab 7. Assignment description at [[BISC_219/F10: Assignment_Series2_DNA Sequencing]]<BR>
	Scientific research report on Series2 Classical (Forward) Genetics Project due week of 11/1 at the beginning of lab. 5%/day late penalty! See assignment directions at: [[BISC 219/F10: Assignment_ Series2_Classical Genetics Paper]]		Scientific research report on Series2 Classical (Forward) Genetics Project due week of 11/1 at the beginning of lab. 5%/day late penalty! See assignment directions at: [[BISC 219/F10: Assignment_ Series2_Classical Genetics Paper]]
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	! 7		! 7
	\| Oct. 20 to <br> Oct. 26		\| Oct. 20 to <br> Oct. 26
-	\| '''Series3:Reverse Genetics''' Colony PCR to look for bacterial transformants	+	\| '''Series2:Finish Mapping:''' SCORE! Calculate Recombination frequency; <BR>'''Series3:Reverse Genetics''' Colony PCR to look for bacterial transformants
-	\| '''Day before lab:''' grow overnight culture of positive colony	+	\| Determine map distance between your mutant gene <BR>and the known reference mutation'''Day before lab:''' grow overnight culture of positive colony
-	\| '''Homework''': ~~Continue working~~ on your Series 2: Forward genetics paper due week of Nov 1	+	\| '''Homework''': Work on your Series 2: Forward genetics paper due week of Nov 1
	\|-		\|-
	! 8		! 8

←Older revision		Revision as of 01:48, 1 October 2010
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	[[BISC 219/F10: Lab 3 \| Lab 3: Linkage Test Part 1]]<br>		[[BISC 219/F10: Lab 3 \| Lab 3: Linkage Test Part 1]]<br>
	[[BISC 219/F10: Lab 4 \| Lab 4: Linkage Test Part 2, Mapping and Complementation]]<br>		[[BISC 219/F10: Lab 4 \| Lab 4: Linkage Test Part 2, Mapping and Complementation]]<br>
-	[[BISC 219/F10: Lab 5 \| Lab 5: Mapping Con't]]<br>	+	[[BISC 219/F10: Lab 5 \| Lab 5: Finish Complementation; Mapping Con't]]<br>
-	[[BISC 219/F10: Lab 6 \| Lab 6: ~~Finish Complementation~~; Mapping Con't]]<BR>	+	[[BISC 219/F10: Lab 6 \| Lab 6: DNA sequence analysis; Mapping Con't]]<BR>
	[[BISC 219/F10: Lab 7 \| Lab 7: Complete Mapping: Score]]<br>		[[BISC 219/F10: Lab 7 \| Lab 7: Complete Mapping: Score]]<br>
	Series3:<BR>		Series3:<BR>